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Cancer Letters Aug 2008We analyzed the mechanism of action for perifosine (D-21266), a new synthetic alkylphospholipid Akt inhibitor, using LNCaP and PC-3 prostate cancer cells. Perifosine...
We analyzed the mechanism of action for perifosine (D-21266), a new synthetic alkylphospholipid Akt inhibitor, using LNCaP and PC-3 prostate cancer cells. Perifosine treatment of PC-3 cells resulted in cytostatic and cytotoxic effects. Cytostatic effects were characterized by cell growth arrest, cell cycle block, and morphological changes, such as a cell enlargement and granulation, hallmarks of differentiating PC-3 cells. Specific differentiation markers including prostasomal, secretory and plasma membrane proteins, and keratins were induced by perifosine. Among them, we detected strong induction and secretion of CEACAM5 protein. In contrast, perifosine strongly reduced caveolin-1 RNA levels. Cytotoxic effects included para-apoptosis, apoptosis, and necrosis. To pursue the mechanisms responsible for these activities we focused on signaling pathways that lie downstream of Akt. Perifosine-triggered GSK-3beta activation in PC-3 and LNCaP cells resulted in the expression of GSK-3beta-related differentiation markers. This expression was reduced in the presence of specific siRNA for GSK-3beta or for its target CREB protein. The use of the GSK-3beta inhibitor lithium chloride indicated that GSK-3beta partially protects prostate cancer cells from the cytotoxic effects of perifosine. Together, these findings indicate that perifosine induces GSK-3beta-related differentiation and caspase-independent cell death in prostate cancer PC-3 cells. In addition our results identify specific biomarkers for perifosine therapy.
Topics: Active Transport, Cell Nucleus; Androgens; Antineoplastic Agents; Apoptosis; Biomarkers; Cell Cycle; Cell Differentiation; Cell Growth Processes; Cell Line, Tumor; Cell Nucleus; Cyclic AMP Response Element-Binding Protein; Gene Expression; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Male; Neoplasms, Hormone-Dependent; Phosphorylcholine; Prostatic Neoplasms; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt
PubMed: 18395973
DOI: 10.1016/j.canlet.2008.02.060 -
Cancer Dec 2011Neuroblastoma (NB) tumors expressing high levels of brain-derived neurotrophic factor (BDNF) and its receptor TrkB or activated Akt are associated with decreased...
BACKGROUND
Neuroblastoma (NB) tumors expressing high levels of brain-derived neurotrophic factor (BDNF) and its receptor TrkB or activated Akt are associated with decreased event-free or overall survival in patients with NB. In the current study, the effect of perifosine, an Akt inhibitor, on the chemosensitivity of TrkB-expressing NB cells or tumors was evaluated.
METHODS
A tetracycline-regulated TrkB-expressing isogenic NB cell model system was tested. In this system, NB cells were treated with etoposide and/or perifosine both in vitro and in vivo. Inhibition of the target by perifosine was evaluated by Western blot analysis or kinase activity assay. Cell survival and tumor growth were investigated.
RESULTS
In vitro BDNF treatment induced Akt phosphorylation and rescued cells from etoposide-induced cell death in cells with high TrkB expression, but not in cells with low TrkB expression. Pretreatment of high TrkB-expressing TB3 cells with perifosine blocked BDNF/TrkB-induced Akt phosphorylation and inhibited BDNF's protection of TB3 cells from etoposide treatment. In vivo, tumors with high TrkB expression were found to have elevated levels of phosphorylated Akt and were less sensitive to etoposide treatment compared with tumors with low TrkB expression. Mice treated with a combination of perifosine and etoposide were found to have a statistically significant decrease in tumor growth compared with mice treated with either etoposide or perifosine alone. Activation of Akt through the BDNF/TrkB signaling pathway induced chemoresistance in NB in vivo.
CONCLUSIONS
Perifosine-induced inhibition of Akt increased the sensitivity of NB to chemotherapy. The results of the current study support the future clinical evaluation of an Akt inhibitor combined with cytotoxic drugs for the improvement of treatment efficacy.
Topics: Brain Neoplasms; Brain-Derived Neurotrophic Factor; Cell Line, Tumor; Drug Resistance, Neoplasm; Etoposide; Humans; Neuroblastoma; Phosphorylation; Phosphorylcholine; Proto-Oncogene Proteins c-akt; Receptor, trkB; Tetracycline
PubMed: 21590687
DOI: 10.1002/cncr.26133 -
Neuroscience Bulletin Apr 2018Accumulating data have revealed that abnormal activity of the mTOR (mammalian target of rapamycin) pathway plays an important role in epileptogenesis triggered by...
Accumulating data have revealed that abnormal activity of the mTOR (mammalian target of rapamycin) pathway plays an important role in epileptogenesis triggered by various factors. We previously reported that pretreatment with perifosine, an inhibitor of Akt (also called protein kinase B), abolishes the rapamycin-induced paradoxical increase of S6 phosphorylation in a rat model induced by kainic acid (KA). Since Akt is an upstream target in the mTOR signaling pathway, we set out to determine whether perifosine has a preventive effect on epileptogenesis. Here, we explored the effect of perifosine on the model of temporal epilepsy induced by KA in rats and found that pretreatment with perifosine had no effect on the severity or duration of the KA-induced status epilepticus. However, perifosine almost completely inhibited the activation of p-Akt and p-S6 both acutely and chronically following the KA-induced status epilepticus. Perifosine pretreatment suppressed the KA-induced neuronal death and mossy fiber sprouting. The frequency of spontaneous seizures was markedly decreased in rats pretreated with perifosine. Accordingly, rats pretreated with perifosine showed mild impairment in cognitive functions. Collectively, this study provides novel evidence in a KA seizure model that perifosine may be a potential drug for use in anti-epileptogenic therapy.
Topics: Animals; Anticonvulsants; Brain; Convulsants; Disease Models, Animal; Epilepsy, Temporal Lobe; Kainic Acid; Male; Neurons; Phosphorylcholine; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Status Epilepticus
PubMed: 28786074
DOI: 10.1007/s12264-017-0165-7 -
Clinical Cancer Research : An Official... Aug 2008The aim of this study is to investigate the efficacy of a novel Akt inhibitor, perifosine, in combination with tumor necrosis factor-related apoptosis-inducing ligand...
PURPOSE
The aim of this study is to investigate the efficacy of a novel Akt inhibitor, perifosine, in combination with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in human myeloma cells and primary patient samples.
EXPERIMENTAL DESIGN
The activity of perifosine in combination with TRAIL was evaluated with experiments testing the effect of perifosine on DR4/DR5 expression by the use of chimeric blocking antibodies, as well as siRNA.
RESULTS
DR4 and DR5 expression was induced by exposure to single-agent perifosine. After exposure of human myeloma cell lines or primary patient samples to increasing doses of perifosine with exogenous TRAIL, we identified synergistically enhanced apoptosis when compared with the perifosine alone, which was achieved with levels well below clinically achievable concentrations for both agents. Transfection with siRNA against DR4, and DR5 reduced the level of apoptosis induced by the combination but did not result in total abrogation of the combination effect. Overexpression of activated Akt, the proposed target for perifosine, did not inhibit the combination effect. Anti-DR4 and DR5 chimeric proteins blocked the cytotoxicity induced by the combination, and the use of c-FLICE-like interleukin protein (FLIP) siRNA enhanced the efficacy at the combination, further supporting the importance of the DR4/DR5 axis in the effect of perifosine.
CONCLUSION
Our observation seems to be independent of the effects of perifosine on Akt signaling, and may represent an additional mechanism of action for this agent, and supports future clinical trials combining these two agents.
Topics: Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Blotting, Western; CASP8 and FADD-Like Apoptosis Regulating Protein; Cell Line, Tumor; Drug Synergism; Flow Cytometry; Humans; Multiple Myeloma; Phosphorylcholine; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Receptors, Death Domain; TNF-Related Apoptosis-Inducing Ligand; Up-Regulation
PubMed: 18698026
DOI: 10.1158/1078-0432.CCR-08-0016 -
PloS One 2012PI3K/AKT signalling pathway is aberrantly active and plays a critical role for cell cycle progression of human malignant pleural mesothelioma (MMe) cells. AKT is one of...
BACKGROUND
PI3K/AKT signalling pathway is aberrantly active and plays a critical role for cell cycle progression of human malignant pleural mesothelioma (MMe) cells. AKT is one of the important cellular targets of perifosine, a novel bio-available alkylphospholipid that has displayed significant anti-proliferative activity in vitro and in vivo in several human tumour model systems and is currently being tested in clinical trials.
METHODS
We tested Perifosine activity on human mesothelial cells and different mesothelioma cell lines, in order to provide evidence of its efficacy as single agent and combined therapy.
RESULTS
We demonstrate here that perifosine, currently being evaluated as an anti-cancer agent in phase 1 and 2 clinical trials, caused a dose-dependent reduction of AKT activation, at concentrations causing MMe cell growth arrest. In this study we firstly describe that MMe cells express aside from AKT1 also AKT3 and that either the myristoylated, constitutively active, forms of the two proteins, abrogated perifosine-mediated cell growth inhibition. Moreover, we describe here a novel mechanism of perifosine that interferes, upstream of AKT, affecting EGFR and MET phosphorylation. Finally, we demonstrate a significant increase in cell toxicity when MMe cells were treated with perifosine in combination with cisplatin.
CONCLUSIONS
This study provides a novel mechanism of action of perifosine, directly inhibiting EGFR/MET-AKT1/3 axis, providing a rationale for a novel translational approach to the treatment of MMe.
Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Cell Line, Tumor; Cisplatin; ErbB Receptors; Humans; Mesothelioma; Phosphorylation; Phosphorylcholine; Pleural Neoplasms; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-met; Signal Transduction
PubMed: 22590625
DOI: 10.1371/journal.pone.0036856 -
Cancer Chemotherapy and Pharmacology Apr 2014The pancreatic cancer has extremely low overall 5-year survival, and gemcitabine is the only approved single agent for pancreatic cancer treatment.
PURPOSE
The pancreatic cancer has extremely low overall 5-year survival, and gemcitabine is the only approved single agent for pancreatic cancer treatment.
METHODS
In the present study, we investigated the potential effect of perifosine, a novel Akt inhibitor on gemcitabine-induced anti-pancreatic cancer effect both in vivo and in vitro.
RESULTS
We showed that sub-cytotoxic low concentration of perifosine dramatically enhanced gemcitabine-induced cytotoxicity in cultured pancreatic cancer cells. Perifosine inhibited Akt-mammalian target of rapamycin and Erk-mitogen-activated protein kinase activation in pancreatic cancer cells. Meanwhile, perifosine suppressed the hedgehog signaling, as it inhibited glioma-associated oncogenes (Gli) 1 activation and decreased its target protein patched 1 (PTCH1) expression. Our data demonstrated that perifosine blocked p70S6K1 (S6K1) activation, thus disrupting S6K1-Gli1 association and subsequent Gli1 activation. The reduction of S6K1 or Gli1 expression by target siRNAs inhibited PTCH1 expression and enhanced gemcitabine-induced cytotoxicity in pancreatic cancer cells. Significantly, perifosine dramatically enhanced gemcitabine-mediated antitumor effect in a PANC-1 xenograft severe combined immunodeficiency mice model.
CONCLUSIONS
In summary, we conclude that perifosine sensitizes gemcitabine-mediated anti-pancreatic cancer efficiency through regulating multiple signaling pathways.
Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Cell Line, Tumor; Deoxycytidine; Disease Progression; Dose-Response Relationship, Drug; Drug Synergism; Gene Knockdown Techniques; Humans; Male; Mice; Mice, SCID; Pancreatic Neoplasms; Phosphorylcholine; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Random Allocation; Ribosomal Protein S6 Kinases, 70-kDa; Signal Transduction; Survival Analysis; Transcription Factors; Xenograft Model Antitumor Assays; Zinc Finger Protein GLI1; Gemcitabine
PubMed: 24519751
DOI: 10.1007/s00280-014-2397-9 -
Cornea May 2010To report a case of unilateral rapidly progressing corneal ring infiltration in a patient being treated with the investigational drug perifosine for Waldenström's...
PURPOSE
To report a case of unilateral rapidly progressing corneal ring infiltration in a patient being treated with the investigational drug perifosine for Waldenström's macroglobulinemia.
METHODS
Interventional case report.
RESULTS
A 55-year-old woman who was receiving treatment with perifosine for Waldenström's macroglobulinemia developed a corneal ring infiltrate in her left eye. She was treated with topical antibiotics and systemic antiviral medication, but the ring infiltrate continued to expand, resulting in complete stromal opacification and a total epithelial defect. Repeated microbiologic tests, including corneal biopsy, failed to identify an infectious agent. Weeks after instituting systemic immunosuppressive medications, including corticosteroids and cyclophosphamide, the ocular inflammation gradually resolved with corneal re-epithelialization and severe stromal neovascularization.
CONCLUSION
Patients treated with perifosine should be monitored for ocular adverse events. Perifosine-related corneal infiltration may progress rapidly and, after appropriate microbiologic examination, aggressive anti-inflammatory therapy should be considered.
Topics: Corneal Opacity; Disease Progression; Female; Humans; Keratoplasty, Penetrating; Middle Aged; Phosphorylcholine; Waldenstrom Macroglobulinemia
PubMed: 20335808
DOI: 10.1097/ICO.0b013e3181b55cd8 -
Tumour Biology : the Journal of the... Jan 2016Rhabdomyosarcoma (RMS) is a highly malignant and metastatic pediatric cancer that arises from the skeletal muscle. Recent studies have identified an important role of...
Rhabdomyosarcoma (RMS) is a highly malignant and metastatic pediatric cancer that arises from the skeletal muscle. Recent studies have identified an important role of AKT signaling in RMS progression. In the current study, we investigated the activity of perifosine, an oral alkylphospholipid AKT inhibitor, against human RMS cells (RD and Rh-30 lines) both in vivo and in vitro, and studied the underlying mechanisms. We showed that perifosine significantly inhibited RMS cell growth in concentration- and time-dependent manners. Meanwhile, perifosine induced dramatic apoptosis in RMS cells. At the signaling level, perifosine blocked AKT activation, while inducing reactive oxygen species (ROS) production as well as JNK and P38 phosphorylations in RMS cells. Restoring AKT activation by introducing a constitutively active-AKT (CA-AKT) only alleviated (not abolished) perifosine-induced cytotoxicity in RD cells. Yet, the ROS scavenger N-acetyl cysteine (NAC) as well as pharmacological inhibitors against JNK (SP-600125) or P38 (SB-203580) suppressed perifosine-induced cytotoxicity in RMS cells. Thus, perifosine induces growth inhibition and apoptosis in RMS cells through mechanisms more than just blocking AKT. In vivo, oral administration of perifosine significantly inhibited growth of Rh-30 xenografts in severe combined immunodeficient (SCID) mice. Our data indicate that perifosine might be further investigated as a promising anti-RMS agent.
Topics: Animals; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Survival; Disease Models, Animal; Drug Evaluation, Preclinical; Humans; MAP Kinase Signaling System; Male; Mice; Mice, SCID; Phosphorylcholine; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Rhabdomyosarcoma; Signal Transduction; Xenograft Model Antitumor Assays
PubMed: 26269112
DOI: 10.1007/s13277-015-3740-4 -
Acta Pharmacologica Sinica Apr 2012The efficacy of the Akt inhibitor perifosine against chronic myeloid leukemia (CML) cells and its mechanisms of action are unknown. In this study, the cytotoxic effects...
AIM
The efficacy of the Akt inhibitor perifosine against chronic myeloid leukemia (CML) cells and its mechanisms of action are unknown. In this study, the cytotoxic effects of perifosine on CML and acute myeloid leukemia (AML) cell lines were compared to elucidate the mechanisms underlying the differences.
METHODS
Human AML cell lines Kasumi-1 and HL-60, and the CML cell line K562 were used. Cell viability was quantitated using MTT assay. Apoptosis was determined using Annexin V-FITC/propidium iodide and Hoechst staining, which were followed by flow cytometry and fluorescence microscopy analysis, respectively. Caspase pathway activation and the expression of autophagy-related genes were examined using Western blot. Autophagy was studied using electron microscopy, the acridine orange staining method, and GFP-LC3 was examined with fluorescence microscopy.
RESULTS
In contrast to AML cell lines, the CML cell lines K562 and K562/G (an imatinib-insensitive CML cell line) were resistant to perifosine (2.5-20 μmol/L) in respect to inhibiting cell growth and inducing apoptosis. Perifosine (2.5, 5, and 10 μmol/L) inhibited Akt and its phosphorylation in AML cells, but not in CML cells. Treatment with perifosine (20 μmol/L) resulted in autophagy in CML cells as shown by the increased formation of acidic vesicular organelles and the accumulation of LC3-II. Treatment of CML cells with perifosine (5, 10, and 20 μmol/L) dose-dependently upregulated AGT5, but not Beclin 1 at the protein level. Furthermore, inhibition of autophagy by chloroquine (40 nmol/L) significantly suppressed the cell growth and induced apoptosis in CML cells treated with perifosine (20 μmol/L).
CONCLUSION
Our results show that CML cell lines were resistant to the Akt inhibitor perifosine in vitro, which is due to perifosine-induced protective autophagy and upregulation of ATG5.
Topics: Antineoplastic Agents; Apoptosis; Autophagy; Autophagy-Related Protein 5; Cell Line, Tumor; Cell Survival; Humans; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; MAP Kinase Kinase 4; Microtubule-Associated Proteins; Phosphorylcholine; Proto-Oncogene Proteins c-akt; Up-Regulation
PubMed: 22407228
DOI: 10.1038/aps.2011.192 -
Molecular Cancer Therapeutics Sep 2009Perifosine is an orally bioavailable alkylphospholipid currently being tested in phase II clinical trials as a potential anticancer drug. In this study, we reveal a...
Perifosine is an orally bioavailable alkylphospholipid currently being tested in phase II clinical trials as a potential anticancer drug. In this study, we reveal a novel mechanism underlying the anticancer activity of perifosine that involves the induction of cyclooxygenase 2 (COX-2) in human cancer cells. Perifosine induced apoptosis and/or cell cycle arrest in several lung and head and neck cancer cell lines. However, the combination of perifosine with low concentrations of celecoxib rendered cells less sensitive to perifosine both in cell culture systems and in lung cancer xenograft models. Subsequently, we examined the effects of perifosine on COX-2 expression and activity in a set of lung and head and neck cancer cell lines, and found that perifosine rapidly and potently increased COX-2 levels and activity, the degrees of which correlated to the abilities of perifosine to inhibit the growth of cancer cells. We also detected increased COX-2 levels in lung cancer xenografts treated with perifosine. Moreover, blockage of COX-2 induction by both antisense and small interfering RNA approaches decreased cell sensitivity to perifosine. Collectively, these data indicate that the activation of COX-2 contributes to the anticancer activity of perifosine, including apoptosis induction and growth arrest. These data are clinically relevant as they suggest that the combination of perifosine and COX-2 inhibitors such as celecoxib, may produce a potential drug contradiction.
Topics: Antineoplastic Agents; Base Sequence; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Celecoxib; Cell Line, Tumor; Cyclooxygenase 2; Drug Screening Assays, Antitumor; Enzyme-Linked Immunosorbent Assay; Gene Silencing; Humans; Immunohistochemistry; Lung Neoplasms; Phosphorylcholine; Pyrazoles; RNA, Small Interfering; Sulfonamides
PubMed: 19755515
DOI: 10.1158/1535-7163.MCT-09-0390